Fig. 2

PKCζ is required for SAHA-induced PLD2 upregulation. a MDA-MB231 cells were transfected with pGL4-PLD2 and pretreated with various inhibitors, PS-PKCζ (50 μM), Rottlerin (10 μM), AG1487 (10 μM), PDTC (50 μM), rapamycin (10 μM), B581 (50 μM), Bay117085 (5 μM), U0126 (20 μM), SP600125 (50 μM), SB203580 (20 μM), LY294002 (20 μM), PP2 (10 μM), and MTM (5 μM), for 30 min, after which they were treated with SAHA (2 μM) for 15 h. The level of luciferase activity was measured as described in “Materials and methods”. MDA-MB 231 cells were co-transfected with pGL4-PLD2 and DN PKCζ (b) or PKCζ siRNA (c) and then treated with SAHA for 15 h. Promoter assays and western blotting were performed. d MDA-MB 231 cells were treated with SAHA (2 μM) for the indicated time and then analyzed by western blot using the indicated antibodies. The intensity of the indicated bands was normalized to the intensity of their respective α-tubulin bands and quantified against each other. Results are representative of at least four independent experiments and shown as the mean ± SEM. N.S (none significant); **p < 0.001